Interleukin-1β-Induced Disruption of Barrier Function in Cultured Human Corneal Epithelial Cells
Interleukin-1β-Induced Disruption of Barrier Function in Cultured Human Corneal Epithelial Cells
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DOI：
10.1167/iovs.08-2606
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发表时间：
2009-02-01
影响因子：
4.4
通讯作者：
Nishida, Teruo
中科院分区：
文献类型：
--
作者：
Kimura, Kazuhiro;Teranishi, Shinichiro;Nishida, Teruo
目的.角膜上皮的屏障功能有助于角膜的稳态，并受到炎症的损害。角膜上皮的粘附连接（AJs）和紧密连接（TJs）对于细胞粘附和屏障功能是必不可少的。我们检测了促炎细胞因子白细胞介素（IL）-1 β对AJs和TJ的影响以及对猴病毒40转化的人角膜上皮（HCE）细胞屏障功能的影响。通过测量跨上皮电阻（TER）评价屏障功能。AJ蛋白E-cadherin和β-catenin、TJ蛋白ZO-1和occludin以及核因子（NF）-κ B B的p65亚基的亚细胞分布通过免疫荧光染色确定。通过免疫印迹分析检测连接蛋白的表达以及NF-κ B抑制蛋白I κ B-α的磷酸化和降解。IL-1 β诱导ZO-1和occludin从邻近HCE细胞的界面消失，而不影响E-钙粘蛋白或β-连环蛋白的定位。它还以浓度和时间依赖性的方式降低HCE细胞的TER。TJ和AJ蛋白的总体丰度不受IL-1 β的影响。IL-1 β诱导I κ B-α的磷酸化和下调以及p65向细胞核的易位。NF-κ B抑制剂姜黄素阻断IL-1 β对TER和ZO-1和occludin亚细胞定位的影响。IL-1 β诱导ZO-1和occludin从HCE细胞的TJ重新分布，从而以依赖于NF-κ B的方式破坏这些细胞的屏障功能。IL-1 β的这些作用可能导致与眼部炎症相关的角膜上皮屏障功能丧失。（Invest Ophthalmol维斯科学。2009;50：597-603）DOI：10.1167/iovs.08-2606
PURPOSE. The barrier function of the corneal epithelium contributes to corneal homeostasis and is impaired by inflammation. Adherens junctions (AJs) and tight junctions (TJs) of the corneal epithelium are essential for cell adhesion and barrier function. We examined the effects of the proinflammatory cytokine interleukin (IL)-1 beta on AJs and TJs as well as on barrier function in simian virus 40-transformed human corneal epithelial (HCE) cells.METHODS. Barrier function was evaluated by measurement of transepithelial electrical resistance (TER). The subcellular distributions of the AJ proteins E-cadherin and beta-catenin, the TJ proteins ZO-1 and occludin, and the p65 subunit of nuclear factor (NF)-kappa B were determined by immunofluorescence staining. The expression of junctional proteins as well as the phosphorylation and degradation of the NF-kappa B-inhibitory protein I kappa B-alpha were examined by immunoblot analysis.RESULTS. IL-1 beta induced the disappearance of ZO-1 and occludin from the interfaces of neighboring HCE cells without affecting the localization of E-cadherin or beta-catenin. It also reduced the TER of HCE cells in a concentration-and time-dependent manner. The overall abundance of TJ and AJ proteins was not affected by IL-1 beta. IL-1 beta induced the phosphorylation and downregulation of I kappa B-alpha as well as the translocation of p65 to the nucleus. The NF-kappa B inhibitor curcumin blocked the effects of IL-1 beta on both TER and the subcellular localization of ZO-1 and occludin.CONCLUSIONS. IL-1 beta induced the redistribution of ZO-1 and occludin from TJs of HCE cells and thereby disrupted the barrier function of these cells in a manner dependent on NF-kappa B. These effects of IL-1 beta may contribute to the loss of corneal epithelial barrier function associated with ocular inflammation. (Invest Ophthalmol Vis Sci. 2009;50:597-603) DOI:10.1167/iovs.08-2606